Exploring and understanding biological and pathological changes are of great significance for early diagnosis and therapy of diseases. Optical sensing and imaging approaches have experienced major progress in this field. Particularly, an emergence of various functional optical nanoprobes has provided enhanced sensitivity, specificity, targeting ability, as well as multiplexing and multimodal capabilities due to improvements in their intrinsic physicochemical and optical properties. However, one of the biggest challenges of conventional optical nanoprobes is their absolute intensity-dependent signal readout, which causes inaccurate sensing and imaging results due to the presence of various analyte-independent factors that can cause fluctuations in their absolute signal intensity. Ratiometric measurements provide built-in self-calibration for signal correction, enabling more sensitive and reliable detection. Optimizing nanoprobe designs with ratiometric strategies can surmount many of the limitations encountered by traditional optical nanoprobes. This review first elaborates upon existing optical nanoprobes that exploit ratiometric measurements for improved sensing and imaging, including fluorescence, surface enhanced Raman scattering (SERS), and photoacoustic nanoprobes. Next, a thorough discussion is provided on design strategies for these nanoprobes, and their potential biomedical applications for targeting specific biomolecule populations (e.g. cancer biomarkers and small molecules with physiological relevance), for imaging the tumor microenvironment (e.g. pH, reactive oxygen species, hypoxia, enzyme and metal ions), as well as for intraoperative image guidance of tumor-resection procedures.

X-rays are widely applied in clinical medical facilities for radiotherapy (RT) and biomedical imaging. However, the sole use of X-rays for cancer treatment leads to insufficient radiation energy deposition due to the low X-ray attenuation coefficients of living tissues and organs, producing unavoidable excessive radiation doses with serious side effects to healthy body parts. Over the past decade, developments in materials science and nanotechnology have led to rapid progress in the field of X-ray-activated tumor-targeting nanosystems, which are able to tackle even systemic tumors and relieve the burden of exposure to large radiation doses. Additionally, novel imaging contrast agents and techniques have also been developed. In comparison with conventional external light sources (e.g., near infrared), the X-ray technique is ideal for the activation of nanosystems for cancer treatment and biomedical imaging applications due to its nearly unlimited penetration depth in living tissues and organisms. In this review, we systematically describe the interaction mechanisms between X-rays and nanosystems, and provide an overview of X-ray-sensitive materials and the recent progress on X-ray-activated nanosystems for cancer-associated theranostic applications.